• 한국미생물·생명공학회지
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• 제31권3호
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• pp.216-223
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• 2003

새롭게 분리된 Bacillus cereus H-1으로부터 크기가 45-kDa인 chitosanase를 정제하여 특성을 파악하였고 1.3-kb의 chitosanase 유전자(choA)를 대장균에 클로닝하여 발현시켰다. H-1의 chitosanase 단백질(ChoA)은 ammonium sulfate 침전과 CM-sephadex칼럼 크로마토그래피에 의해 정제하였다. 최적 pH는 약 7이었고 pH 안정성은 $50^{\circ}C$에서 4-11로 나타났다. 최적 온도는 약 5$0^{\circ}C$였으며 효소 활성은 $45^{\circ}C$ 아래에서 비교적 안정하였다. H-1 chitosanase는 soluble 또는 glycol chitosan뿐만아니라 carboxymethyl cellulose(CMC)에 대한 활성도 나타내었다. 정제된 ChoA의 MALDI-TOF MS분석에 기초하여 이미 알려진 다른 Bacillus chitosanases와의 데이터베이스 검색을 통해 전체 아미노산 서열을 밝혀내었다. Chitosanase gene에 해당하는 1.6 kb의 PCR 산물을 얻었으며 그의 DNA 서열을 결정하였다. choA의 추정 아미노산은 Bacillus sp. No 7-M과 Bacillus sp. KCTC0377BP의 아미노산과 98%의 유사성을 나타내었다. 재조합 ChoA단백질은 E. coli DH5$\alpha$에서 원 균주와 동일한 크기로 발현되었다. N말단의 추정아미노산서열을 다른 chitosanas리 서열과 비교해 볼때 ChoA는 chitosanase-cellulase 활성을 갖는 family 8에 속하는 미생물 endo-chitosanaseT. 추정되었다.

• 한국식품저장유통학회지
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• 제19권5호
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• pp.757-766
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• 2012

향미가 우수한 100년 묵은 재래 간장에서 부터 강력한 항진균 활성(antifungal)균주를 분리 동정하여 Bacillus velelzensis SSH100-10으로 명명하였다. B. velezensis SSH100-10은 식품 위해 및 식품 변패곰팡이에 대한 항곰팡이 활성과 장류에 이취를 주는 산막효모 등에 대한 항효모 활성이 동시에 있으며, 강력한 단백분해활성을 나타내었다. NaCl 내염성도 우수하여 12% NaCl 농도하에서도 생육 48시간에 $A_{600}$에서 3.51의 생육도를 나타내었다. 또한 최근 Bacillus subtilis group 중 일부 균주에서 설사형 독소인 enterotixon을 나타내는 균주들에 대한 보고가 증가함에 따라, 안전성검증의 일환으로 enterotoxin 생성 여부를 조사하였을 때, 본 분리 균주는 enterotoxin 검지반응에서 음성을 나타내었다. B. velezensis SSH100-10이 생산하는 항진균 활성물질을 SPE, preparative HPLC, reverse phase-HPLC로 정제 후, MALDI-TOF-MS와 아미노산 조성 분석을 통하여 그 항진균 원인 물질이 $C_{14}$ iturin A와 $C_{15}$ iturin A 임을 구명하였다. 또한 B. velezensis SSH100-10의 배양상징액과 분리 정제된 항진균 물질 $C_{14}$ iturin A와 $C_{15}$ iturin A와 대조구로 시판되는 $C_{14{\sim}15}$ iturin A의 pH, 온도, 효소 안정성 실험을 통하여 B. velezensis SSH100-10은 pH, 열, 효소처리에 매우 안정한 iturin A 이외에도 pH에 불안정한 또 다른 구명되지 않은 항진균 물질을 생산함을 보고하였다. 본 연구에서 보고된 강력한 항진균 활성을 지니는 B. velezensis SSH100-10은 맛있는 발효과정의 종균으로서의 작용과 더불어 콩발효식품에서 유해요소가 될 수 있는 바람직하지 못한 미생물을 제어함으로서 콩발효식품의 안전과 위생수준을 개선할 수 있는 우수한 종균으로 활용될 수 있을 것이다.

• 공업화학
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• 제23권6호
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• pp.529-533
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• 2012

본 연구에서는 온순조건에서 용매 추출된 석탄의 물성과 추출특성을 고온조건에서 추출된 석탄의 것과 비교하였다. 석탄의 용매 추출 특성을 알아보기 위해 추출실험에는 아역청탄(Kideco 탄)과 극성 용매인 N-methyl-2-pyrrolidinone (NMP)을 사용하였다. 온순조건과 고온조건에서 석탄의 용매 추출 특성을 알아보기 위해 추출온도 변화, 용매 재사용에 따른 추출 특성 변화에 대한 실험을 하였다. 실험결과 추출온도가 증가할수록 추출수율과 추출탄의 발열량은 증가 하였고, 회분농도는 감소하였다. FT-IR 분석 결과 고온조건($350^{\circ}C$)에서 추출한 추출탄의 표면에 C=O 아미드 결합, 방향족 에테르, 그리고 지방족 에테르 그룹들이 온순조건($150{\sim}300^{\circ}C$)에서 추출한 것에 비해 증가하는 것으로 나타났다. MALDI-TOF/MS 분석 결과 온순조건에서는 300~500 m/z 범위 이하의 작은 분자들이 주로 추출되었고, 고온조건에서는 500~1500 m/z 범위에 걸쳐 상대적으로 큰 분자들까지 추출됨을 확인하였다.

• 한국환경보건학회지
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• 제50권2호
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• pp.93-101
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• 2024

Background: As pollutants caused by non-point sources flow into rivers, river water quality monitoring for fecal pollution is becoming increasingly important. Objectives: This study was conducted to investigate the distribution of microbial communities in the Yeongsangang River water system and sewage treatment plants in Gwangju and to evaluate their antibiotic resistance. Methods: In the experiment, samples were distributed to five selective media at each point and then cultured for 18 to 24 hours. When bacteria were observed, they were sub-cultured by size and shape and identified using MALDI-TOF MS equipment. When identification was completed, 17 types of antibiotic susceptibility tests were performed using VITEK II equipment, focusing on gram-negative dominant species among the identified strains. Results: During the study period, a total of 266 strains were isolated from 39 samples. Gram-positive bacteria were 37 strains in four genera, or 13.9% of the total, and Gram-negative bacteria were 229 strains in 23 genera, or 86.1% of the total. Antibiotic susceptibility testing of 23 strains, the major dominant species, showed that one strain (4.3%) was resistant to only one antibiotic, and two strains (8.7%) were 100% susceptible to the 17 antibiotics tested. The other 20 strains (87.0%) were multidrug resistant bacteria resistant to two or more antibiotics. There were various types of multidrug resistance. Among them, penicillin and cephalosporin series showed the highest resistance. Conclusions: Based on the results of this study, it was found that the bacterial community structure changed according to regional and environmental factors, and it was judged that continuous research such as genetic analysis of antibiotic-resistant bacteria present in natural rivers is necessary.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1990-1994
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• 2006

Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited a high GA productivity. A gas chromatography-mass spectrometry (GC-MS) analysis of extracts of the culture fluid of F proliferatum KGL0401 also revealed the presence of $GA_1$, $GA_3$, $GA_4$, $GA_7$, $GA_{20}$, and $GA_{24}$. Therefore, the present study conducted a proteome analysis of waito-c rice treated with the culture fluid of the isolated F proliferatum KGL0401 to identify the protein expression triggered by the GA-containing culture fluid. The results revealed the overexpression of 180 protein spots in the sample treated with the culture fluid. Among them, 75 induced proteins were selected and analyzed by MALDI-TOF (matrix-assisted laser desorption-iorrization time-of-flight) mass spectrometry, followed by database searching, and 51 proteins were identified.

• Molecules and Cells
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• 제22권1호
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• pp.36-43
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• 2006

Mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering, but their application has been impeded by lack of knowledge of their core biological properties. In order to identify MSC-specific proteins, the hydrophobic protein fraction was individually prepared from two different umbilical cord blood (UCB)-derived MSC populations; these were then subjected to two-dimensional (2D) gel electrophoresis and peptide mass fingerprinting matrix-assisted laser desorption/ionization (MALDI)-time of flight (TOF)-mass spectrometry (MS). Although the 2D gel patterns differed somewhat between the two samples, computer-assisted image analysis identified shared protein spots. 35 spots were reliably identified corresponding to 32 different proteins, many of which were chaperones. Based on their primary sub-cellular locations the proteins could be grouped into 6 categories: extracellular, cell surface, endoplasmic reticular, mitochondrial, cytoplasmic and cytoskeletal proteins. This map of the water-insoluble proteome may provide valuable insights into the biology of the cell surface and other compartments of human MSCs.

• KSBB Journal
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• 제21권1호
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• pp.16-19
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• 2006

최근의 연구에 의하면 열이나, 산화적 스트레스에 대해서 NDPK는 구조적인 변화를 유발하며, 효소 활성과 구조가 oxidant에 의해 변화된다는 보고를 근거로 하여 정상적인 효모균주와 효모의 NDPK 유전자가 파괴된 mutant에서, 산화적 스트레스에 관련된 역할을 규명하고자 2-D 전기영동 방법을 통해서, $H_2O_2$의 처리전과 처리 후에 전사패턴이 변화된 유전자들, 즉, 산화적조절 신호체제에 연관되어졌을 것이라고 생각되어지는 몇 개의 단백질 리스트를 얻었다. 이 결과는 NDPK의 redox state의 조절에 관련된 효소의 성질을 규명함에 있어 유용한 유전자 신호 체제정보를 제공할 것으로 생각되어진다.

• Parasites, Hosts and Diseases
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• 제55권1호
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• pp.15-20
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• 2017

The aim of this study was to identify antigens for a vaccine or drug target to control rabbit coccidiosis. A combination of 2-dimensional electrophoresis, immunoblotting, and mass spectrometric analysis were used to identify novel antigens from the sporozoites of Eimeria stiedae. Protein spots were recognized by the sera of New Zealand rabbits infected artificially with E. stiedae. The proteins were characterized by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) analysis in combination with bioinformatics. Approximately 868 protein spots were detected by silver-staining, and a total of 41 immunoreactive protein spots were recognized by anti-E. stiedae sera. Finally, 23 protein spots were successfully identified. The proteins such as heat shock protein 70 and aspartyl protease may have potential as immunodiagnostic or vaccine antigens. The immunoreactive proteins were found to possess a wide range of biological functions. This study is the first to report the proteins recognized by sera of infected rabbits with E. stiedae, which might be helpful in identifying potential targets for vaccine development to control rabbit coccidiosis.

• BMB Reports
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• 제38권5호
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• pp.545-549
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• 2005

Acidithiobacillus ferrooxidans is one of the most important bacterium used in bioleaching, and can utilize $Fe^{2+}$ or sulphide as energy source. Growth curves for Acidithiobacillus ferrooxidans under phosphate starvation and normal condition have been tested, showing lag, logarithmic, stationary and aging phases as seen in other bacteria. The logarithmic phases were from 10 to 32 hours for Acidithiobacillus ferrooxidans cultivated with normal cultivating condition and from 20 to 60 hrs for Acidithiobacillus ferrooxidans cultivated phosphate starvation. Differences of protein patterns of Acidithiobacillus ferrooxidans growing in case of normal or phosphate starvation were separately investigated after cultivation at $30^{\circ}C$ by the analysis of two-dimensional gel electrophoresis (2-DE), matrix-assisted laser desorption/ionization (MALDI)-Mass spectrometry. There were total 6 protein spots identified, which were Recombination protein recA, RNA helicase, AP2 domain-containing transcription factor, NADH dehydrogenase I chain D, Hyothetical protein PF1669, and Transaldolase STY3758. From the 6 identified protein spots, 3 proteins were found to be decreased in expression at the cultivating condition of phosphate starvation, while another three upregulated.

• 한국식품영양학회지
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• 제24권3호
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• pp.291-294
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• 2011

A lipase inhibitor from Desmodium oxyphyllum DC. was purified by methanol extraction, systematic solvent extraction, silica gel column chromatography, $C_{18}$ solid phase extraction chromatography and RP-HPLC. We obtained the purified lipase inhibitor with 182 ng($IC_{50}$) of lipase inhibitory activity for a 0.06% yield. Its molecular weight was estimated to be 655.37 Da from an instrumental analysis of MALDI-TOF-MS and it was identified copper-3,5-dibromo-2-hydroxybenzoic acid ($C_{14}H_8Br_4CuO_6$) by $^1H$, $^{13}C$ NMR analysis.